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Identification of new AAV vectors with enhanced blood-brain barrier penetration efficiency via organ-on-a-chip.
Li, Mengmeng; Zhong, Yujian; Zhu, Mingyang; Pang, Chunjin; Xiao, Lu; Bu, Ye; Li, Huapeng; Diao, Yong; Yang, Chaoyong; Liu, Dan.
Afiliación
  • Li M; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
  • Zhong Y; PackGene Biotech, Guangzhou, Guangdong, China.
  • Zhu M; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
  • Pang C; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
  • Xiao L; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
  • Bu Y; PackGene Biotech, Guangzhou, Guangdong, China.
  • Li H; PackGene Biotech, Guangzhou, Guangdong, China.
  • Diao Y; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
  • Yang C; State Key Laboratory of Physical Chemistry of Solid Surfaces, the Key Laboratory of Chemical Biology of Fujian Province, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian, China.
  • Liu D; Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Xiamen, Fujian, China. liudan@hqu.edu.cn.
Analyst ; 149(15): 3980-3988, 2024 Jul 22.
Article en En | MEDLINE | ID: mdl-38872436
ABSTRACT
To overcome limitations in the generalizability and efficiency of current AAV vectors, in this current study, we constructed an AAV variant library by the insertion of random heptapeptide sequences in the receptor-binding domain of the AAV9 capsid gene. We then applied a recently developed organ-on-a-chip in vitro model of the human blood-brain barrier (BBB) to iteratively enrich for variants that efficiently cross the BBB and transduce astrocyte cells. Through multiple rounds of screening, we obtained two candidate AAV variants, AAV-M6 and AAV-M8, which showed significantly higher BBB penetration efficiency than AAV9 or AAV-PHP.eB. Quantitative PCR (qPCR) assay showed that AAV-M6 could accumulate to a 5 times higher titer, while AAV-M8 reached a 3 times higher titer, than AAV-PHP.eB in the neural chamber of the model. The transduction assay further verified that the AAV-M6 candidate vector was able to infect HA-1800 cells after crossing the BBB, suggesting it could potentially transduce brain parenchymal cells after crossing the hCMEC/D3 layer at higher efficiency than AAV-PHP.eB. Molecular simulations suggested that the human receptor proteins, LY6D and M6PR, could bind the AAV-M6 heptapeptide insertion with high affinity. This study provides two promising candidate AAV vectors and demonstrates the use of this in vitro BBB model for scalable, high-throughput screening of gene therapies. These tools can drive investigations of the mechanisms underlying BBB permeability and the cell-type specificity of virus vectors.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Barrera Hematoencefálica / Dependovirus / Vectores Genéticos Límite: Humans Idioma: En Revista: Analyst Año: 2024 Tipo del documento: Article País de afiliación: China

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Barrera Hematoencefálica / Dependovirus / Vectores Genéticos Límite: Humans Idioma: En Revista: Analyst Año: 2024 Tipo del documento: Article País de afiliación: China